Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library
Abstract
Recombinant EstE1 protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was then crystallized at 290 K by the hanging-drop vapour-diffusion method. EstE1, a new thermostable esterase, was isolated by functional screening of a metagenomic DNA library from thermal environment samples. This enzyme showed activity towards short-chain acyl derivatives of length C4–C6 at a temperature of 303–363 K and displayed a high thermostability above 353 K. EstE1 has 64 and 57% amino-acid sequence similarity to est{sub pc}-encoded carboxylesterase from Pyrobaculum calidifontis and AFEST from Archaeoglobus fulgidus, respectively. The recombinant protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was crystallized at 290 K by the hanging-drop vapour-diffusion method. X-ray diffraction data were collected to 2.3 Å resolution from an EstE1 crystal; the crystal belongs to space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 73.71, c = 234.23 Å. Assuming the presence of four molecules in the asymmetric unit, the Matthews coefficient V{sub M} is calculated to be 2.2 Å{sup 3} Da{sup −1} and the solvent content is 44.1%.
- Authors:
- Department of Biology, Yonsei University, Seoul 120-749 (Korea, Republic of)
- (Korea, Republic of)
- Department of Biotechnology, Yonsei University, Seoul 120-749 (Korea, Republic of)
- Publication Date:
- OSTI Identifier:
- 22356264
- Resource Type:
- Journal Article
- Resource Relation:
- Journal Name: Acta Crystallographica. Section F; Journal Volume: 62; Journal Issue: Pt 2; Other Information: PMCID: PMC2150951; PMID: 16511287; PUBLISHER-ID: fw5062; OAI: oai:pubmedcentral.nih.gov:2150951; Copyright (c) International Union of Crystallography 2006; Country of input: International Atomic Energy Agency (IAEA)
- Country of Publication:
- United Kingdom
- Language:
- English
- Subject:
- 75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; AFFINITY; CRYSTALLIZATION; CRYSTALS; DIFFUSION; ENVIRONMENT; ESCHERICHIA COLI; HISTIDINE; IRON; MOLECULES; RESOLUTION; SCREENING; SPACE GROUPS; STRAINS; X-RAY DIFFRACTION
Citation Formats
Byun, Jung-Sue, Protein Network Research Center, Yonsei University, Seoul 120-749, Rhee, Jin-Kyu, Kim, Dong-Uk, Oh, Jong-Won, Cho, Hyun-Soo, E-mail: hscho8@yonsei.ac.kr, and Protein Network Research Center, Yonsei University, Seoul 120-749. Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library. United Kingdom: N. p., 2006.
Web. doi:10.1107/S1744309106000832.
Byun, Jung-Sue, Protein Network Research Center, Yonsei University, Seoul 120-749, Rhee, Jin-Kyu, Kim, Dong-Uk, Oh, Jong-Won, Cho, Hyun-Soo, E-mail: hscho8@yonsei.ac.kr, & Protein Network Research Center, Yonsei University, Seoul 120-749. Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library. United Kingdom. doi:10.1107/S1744309106000832.
Byun, Jung-Sue, Protein Network Research Center, Yonsei University, Seoul 120-749, Rhee, Jin-Kyu, Kim, Dong-Uk, Oh, Jong-Won, Cho, Hyun-Soo, E-mail: hscho8@yonsei.ac.kr, and Protein Network Research Center, Yonsei University, Seoul 120-749. Wed .
"Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library". United Kingdom.
doi:10.1107/S1744309106000832.
@article{osti_22356264,
title = {Crystallization and preliminary X-ray crystallographic analysis of EstE1, a new and thermostable esterase cloned from a metagenomic library},
author = {Byun, Jung-Sue and Protein Network Research Center, Yonsei University, Seoul 120-749 and Rhee, Jin-Kyu and Kim, Dong-Uk and Oh, Jong-Won and Cho, Hyun-Soo, E-mail: hscho8@yonsei.ac.kr and Protein Network Research Center, Yonsei University, Seoul 120-749},
abstractNote = {Recombinant EstE1 protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was then crystallized at 290 K by the hanging-drop vapour-diffusion method. EstE1, a new thermostable esterase, was isolated by functional screening of a metagenomic DNA library from thermal environment samples. This enzyme showed activity towards short-chain acyl derivatives of length C4–C6 at a temperature of 303–363 K and displayed a high thermostability above 353 K. EstE1 has 64 and 57% amino-acid sequence similarity to est{sub pc}-encoded carboxylesterase from Pyrobaculum calidifontis and AFEST from Archaeoglobus fulgidus, respectively. The recombinant protein with a histidine tag at the C-terminus was overexpressed in Escherichia coli strain BL21(DE3) and then purified by affinity chromatography. The protein was crystallized at 290 K by the hanging-drop vapour-diffusion method. X-ray diffraction data were collected to 2.3 Å resolution from an EstE1 crystal; the crystal belongs to space group P4{sub 1}2{sub 1}2, with unit-cell parameters a = b = 73.71, c = 234.23 Å. Assuming the presence of four molecules in the asymmetric unit, the Matthews coefficient V{sub M} is calculated to be 2.2 Å{sup 3} Da{sup −1} and the solvent content is 44.1%.},
doi = {10.1107/S1744309106000832},
journal = {Acta Crystallographica. Section F},
number = Pt 2,
volume = 62,
place = {United Kingdom},
year = {Wed Feb 01 00:00:00 EST 2006},
month = {Wed Feb 01 00:00:00 EST 2006}
}
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The catalytic domain of the glucuronoyl esterase from H. jecorina was overexpresssed, purified and crystallized in space group P2{sub 1}2{sub 1}2{sub 1}. X-ray diffraction data were collected to 1.9 Å resolution. The catalytic domain of the glucuronoyl esterase from Hypocrea jecorina (anamorph Trichoderma reesei) was overexpresssed, purified and crystallized by the sitting-drop vapor-diffusion method using 1.4 M sodium/potassium phosphate pH 6.9. The crystals belonged to space group P2{sub 1}2{sub 1}2{sub 1} and X-ray diffraction data were collected to 1.9 Å resolution. This is the first enzyme with glucoronoyl esterase activity to be crystallized; its structure will be valuable in lignocellulose-degradationmore »